S. epidermidis (ATCC 35984) was obtained from American Type Culture Collection. Bacteria were grown overnight at 37 °C on tryptic soy agar (TSA), (Merck, Kenilworth, NJ, USA) or on tryptic soy broth (TSB), (Merck, Kenilworth, NJ, USA) at 150 rpm. Inoculum was adjusted by measuring optical density (OD) at 600 nm and confirmed by colony-forming unit (CFU) counts.
S epidermidis atcc 35984
Manufactured by American Type Culture Collection
Sourced in United States
S. epidermidis ATCC 35984 is a strain of Staphylococcus epidermidis, a Gram-positive bacterium. It is a reference strain available from the American Type Culture Collection (ATCC) for research and laboratory applications.
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Lab products found in correlation
S epidermidis atcc 12228, by American Type Culture Collection (2 mentions)
S aureus atcc 43300, by American Type Culture Collection (2 mentions)
S aureus atcc 29213, by American Type Culture Collection (2 mentions)
Tobramycin, by Merck Group (1 mentions)
Vancomycin, by Merck Group (1 mentions)
Merck silica gel 60 f254, by Merck Group (1 mentions)
Agilent lc ms esi tof 1260 6230da, by Agilent Technologies (1 mentions)
Imipenem, by Merck Group (1 mentions)
Oxacillin, by Merck Group (1 mentions)
Pseudomonas aeruginosa atcc 27853, by American Type Culture Collection (1 mentions)
Staphylococcus aureus atcc 43300, by American Type Culture Collection (1 mentions)
Ethanol, by Merck Group (1 mentions)
Nutrient broth, by Thermo Fisher Scientific (1 mentions)
Tryptone soya agar, by Thermo Fisher Scientific (1 mentions)
1 2 distearoyl sn glycero 3 phosphocholine dspc, by Lipoid (1 mentions)
Mpeg2000 dspe, by Lipoid (1 mentions)
Bacteriological agar, by Thermo Fisher Scientific (1 mentions)
S aureus atcc 6538, by American Type Culture Collection (1 mentions)
Mrsa mu50, by American Type Culture Collection (1 mentions)
Tryptic soya broth tsb, by Thermo Fisher Scientific (1 mentions)
7 protocols using s epidermidis atcc 35984
1
Cultivation and Quantification of S. epidermidis
2
Analytical Characterization of Antimicrobial Compounds
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1 H and 1 3C-NMR spectra were recorded at 400/100 or 500/125 MHz in CDCl 3 or CD 3 OD on Bruker (Bremen, Germany) or INOVA 500 (Palo Alto, CA, USA) spectrometers. LC-MS analyses were run on an Agilent LC-MS ESI-TOF 1260/ 6230DA (Cernusco sul Naviglio, Milan, Italy) instrument operating in positive ionization mode. Analytical TLC was performed on Merck silica gel (60F254) precoated plates (0.20 m). The compounds were visualized under UV light (254 nm). Column chromatography was performed on silica gel with pore size 60 Å, 60-400 mesh particle size with the indicated solvents.
All the reagents were purchased from Sigma-Aldrich (Milan, Italy) and used without any further purification. Vancomycin, oxacillin, tobramycin, and imipenem were purchased from Sigma-Aldrich (Milan, Italy) . Staphylococcus aureus ATCC 43300 (MRSA), S. epidermidis ATCC 29213, S. epidermidis ATCC 35984, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC BAA-1705 (carbapenemase producer), were obtained from the American Type Culture Collection (Rockville, MD).
All the reagents were purchased from Sigma-Aldrich (Milan, Italy) and used without any further purification. Vancomycin, oxacillin, tobramycin, and imipenem were purchased from Sigma-Aldrich (Milan, Italy) . Staphylococcus aureus ATCC 43300 (MRSA), S. epidermidis ATCC 29213, S. epidermidis ATCC 35984, Pseudomonas aeruginosa ATCC 27853, Klebsiella pneumoniae ATCC BAA-1705 (carbapenemase producer), were obtained from the American Type Culture Collection (Rockville, MD).
3
Antimicrobial Activity Assessment
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S. aureus ATCC 29213, S. aureus ATCC 43300, S. epidermidis ATCC 12228 and S. epidermidis ATCC 35984, were obtained from the American Type Culture Collection (Rockville, MD, USA). All the strains were cultured in tryptic soya broth (TSB, OXOID) under aerobic conditions at 37 °C for 24 h on an orbital shaker at 200 rpm. Each tested compound was dissolved in ethanol (Sigma, Milan, Italy) and diluted in TSB to give a stock solution.
4
Microbial Colonization on Hernia Meshes
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S. aureus ATCC 6538, S. aureus ATCC 700699 (also known as MRSA Mu50) and S. epidermidis ATCC 35984 were purchased from the American Type Culture Collection (Manassas, VA, USA). Bacteria were inoculated at colony forming unit (CFU)/ml or optical density at 600 nm (OD600) values stated after dilution of an overnight culture grown in tryptone soya broth (TSB) or nutrient broth (Thermo Fisher Scientific, Waltham, MA, USA) at 37 °C with shaking at 180 rpm. Tryptone soya agar (TSA) was prepared by adding 1.5% agar bacteriological (Thermo Fisher Scientific). The hernia meshes Parietex Hydrophilic 2-Dimensional mesh (polyester), Parietene Lightweight monofilament polypropylene mesh (polypropylene) were donated by Covidien (Dublin, Ireland). The saturated phospholipids 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy (polyethylene glycerol)-2000] (DSPE-mPEG2000) were donated by Lipoid GmbH (Ludwigshafen, Germany). Unless stated otherwise, all chemicals, materials, media and supplements were purchased from Sigma-Aldrich (Steinheim, Germany).
5
Staphylococcus Biofilm Formation Protocols
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All of the bacterial strains and plasmids used in the study are shown in Table 4 . The biofilm-positive strain S. epidermidis ATCC 35984 (RP62A; GenBank accession number NC_002976 ) and non-biofilm-forming strain ATCC 12228 (GenBank accession number NC_004461 ) were purchased from the American Type Culture Collection (ATCC; Manassas, VA) (50 (link)). S. epidermidis 1457 (SE1457) and S. aureus RN4220 were provided by Gao Fu from the University of Hong Kong. The S. epidermidis strains and S. aureus RN4220 were cultured in tryptic soya broth (TSB; Oxoid, Basingstoke, UK) at 37°C with shaking at 220 rpm. Glucose was added to the TSB medium at a concentration of 0.5% for the detection of biofilm formation. Electroporation was used for plasmid transformation, and B2 medium (1% casein hydrolysate, 2.5% yeast extract, 0.5% glucose, 2.5% NaCl, 0.1% K2HPO4, pH 7.5) was used for the recovery of bacteria. The antibiotics used in this study were purchased from Sigma Chemical Co. (Los Angeles, CA, USA) and used at concentrations of 10 mg/liter for chloramphenicol, 100 mg/liter for ampicillin, 50 ng/ml for anhydrotetracycline, and 10 mg/liter for erythromycin.
6
Staphylococcus Isolates for Anti-Microbial Screening
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All growth media and chemicals were purchased from Sigma-Aldrich (UK) unless otherwise stated. S. aureus DSMZ11729 (ATCC 33592) was obtained from the Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH (Germany). S. aureus ATCC 25923, S. aureus BAA-1717, S. epidermidis ATCC 35984, and S. epidermidis ATCC 12228 were obtained from the American Type Culture Collection (LGC Standards, UK). S. aureus NCTC10442, S. aureus NCTC10788 (ATCC 6538), and S. aureus NCTC6571 (ATCC 9144) were obtained from the National Collection of Type Cultures (Public Health England, UK). S. aureus SMRSA105, S. aureus SMRSA124, S. aureus SMRSA161, S. aureus EMRSA16, S. aureus MRSA8, S. aureus MRSA10, S. aureus MSSA1, S. aureus MSSA2, S. aureus MSSA6, and S. aureus MSSA8 were all isolated from dermal wounds and were generous gifts from Gail Ferguson and Isabel Cook of the Medical Microbiology Department, Aberdeen Royal Infirmary (UK). All other isolates were from the anterior nares of healthy volunteers.
7
Characterization of S. aureus Isolates
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A total of three hundred (n=300) S. aureus isolates (corresponding to MSSA and MRSA isolates in equal measure; n=150 isolates, respectively) were included in this study, which were kindly provided by a tertiary-care teaching hospital and two smaller regional hospitals in Hungary. The study uses a cross-sectional study design; the microorganisms were isolated between 2019.01.01 and 2020.01.01., including n=100 isolates from catheter-associated infections (CAI-SA), skin and soft tissue infections (SSTI-SA) and urinary tract infections (UTI-SA). During our experiments, S. aureus ATCC 29213 (MSSA; positive for biofilm-production, icaAB gene negative), S. aureus ATCC 43300 (MRSA; positive for biofilm-production, icaAB gene positive), S. aureus ATCC 12600 (MSSA; non-biofilm producing, icaAB gene negative), S. epidermidis ATCC 35984 (positive for biofilm-production, icaAB gene positive) and S. epidermidis ATCC 12224 (non-biofilm producing, icaAB gene negative) were used as control strains, obtained from the American Type Culture Collection (ATCC; Manassas, VI, USA).39 Stock cultures were stored at −80 °C in a cryopreservation medium (700 µL trypticase soy broth + 300 µL 50% glycerol).
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